Effects of kinins on isolated stomachs of control and transgenic knockout B2 receptor mice.

Abstract

The aim of this study was to investigate the pharmacological profile of the kinin B1 and B2 receptors in isolated stomachs from wild-type control and B2 receptor knockout mice. Isometric contractions evoked by bradykinin (BK) (9 nM) and desArg9BK (28 nM) were shown to be different. The contraction induced by desArg9BK had a longer duration than that evoked by BK and increased during incubation in vitro in stomachs of wild-type controls, while in the transgenic B2 receptor knockout mice, the contractions evoked by desArg9BK and BK were similar and followed the B1 receptor agonist pattern. BK but not the carboxypeptidase-resistant analog, [Phe8psi(CH2-NH)Arg9]BK, was found to be active in the stomach of B2 receptor knockout mice. BK-induced contractions were prevented by mergetpa (a carboxypeptidase M inhibitor) (10 microM) and by a the B receptor antagonist, AcLys[DbetaNal7,Ile8]desArg9BK (R 715) (0.88 microM), while not being influenced by the B2 receptor antagonist HOE 140 (0.38 microM). BK and [Phe8psi(CH2-NH)Arg9]BK were potent contractants of the wild-type mice stomach and their effects were not influenced by mergetpa or by the B receptor antagonist: they were reduced by HOE 140. After incubation in vitro for 3-4 hours, the tissues were treated with HOE 140 (4 microM) and FR-173657 (17 microM) to eliminate B2 receptor function. In these tissues, BK evoked a B1-like contraction which was inhibited by mergetpa (10 microM) and antagonized by R 715 (8 microM). The results indicate that BK acts primarily on B2 receptors. However, after intramural conversion to desArg9BK, activation of B1 receptors of the mice stomach occurs. In the tissues of B2 receptor knockout mice, BK behaves as a pure B1 receptor agonist while in stomachs of control animals, the B2 receptor contribution is overwhelming. After complete blockade of the B2 receptor, BK is able to evoke B1-mediated responses similar to those observed in tissues of B2 receptor knockout mice. It is concluded that the disruption of the B2 receptor gene eliminates the B2 receptor without influencing the B1 receptor system.